Manish Kumar Gautam

Evaluation of in vivo wound healing activity of Bacopa monniera on different wound model in rats.

Wound healing effects of 50% ethanol extract of dried whole plant of Bacopa monniera (BME) was studied on wound models in rats. BME (25 mg/kg) was administered orally, once daily for 10 days (incision and dead space wound models) or for 21 days or more (excision wound model) in rats. BME was studied for its in vitro antimicrobial and in vivo wound breaking strength, WBS (incision model), rate of contraction, period of epithelization, histology of skin (excision model), granulation tissue free radicals (nitric oxide and lipid peroxidation), antioxidants (catalase, superoxide dismutase, and reduced glutathione), acute inflammatory marker (myeloperoxidase), connective tissue markers (hydroxyproline, hexosamine, and hexuronic acid), and deep connective tissue histology (dead space wound). BME showed antimicrobial activity against skin pathogens, enhanced WBS, rate of contraction, skin collagen tissue formation, and early epithelization period with low scar area indicating enhanced healing. Healing effect was further substantiated by decreased free radicals and myeloperoxidase and enhanced antioxidants and connective tissue markers with histological evidence of more collagen formation in skin and deeper connective tissues. BME decreased myeloperoxidase and free radical generated tissue damage, promoting antioxidant status, faster collagen deposition, other connective tissue constituent formation, and antibacterial activity.

In–vitro antibacterial activity on human pathogens and total phenolic, flavonoid contents of Murraya paniculata Linn. leaves

Abstract Objective To deals with in–vitro antibacterial analysis of Murraya paniculata Linn. (Rutaceae) leaves extract on human pathogens in different solvent and determination of total phenolic and flavonoid contents in petroleum ether, methanol, ethanol and hydro-alcoholic extract of the plant. Methods These extracts were tested against various human pathogens for antimicrobial activity which was evaluated by disc diffusion method and minimum inhibitory concentration (MIC) and minimum bactericidal concentration was calculated by micro dilution method. Phenolic content was estimated by using folin ciocalteau reagent and flavonoids by using aluminium chloride reagent against quercetin equivalent. Results The methanolic extract of leaf showed marked antibacterial activities against gram-positive and gram-negative bacteria. Methanolic extract of Murraya paniculata leaf contain higher phenolic content (24.80±0.64) followed by ethanolic fraction (15.40±0.38), pet. Ether (13.50±0.96) and hydro-alcoholic (9.06±1.13). Flavonoid content was found to be maximum in pet. ether extracts (3.38±1.89). Conclusions Murraya paniculata leaves posses antibacterial properties against human pathogens with high content of phenolic and flavonoids, which have supportive action of antibacterial activity. Studying plant based antimicrobial properties provides additional information in developing nature antibiotics and discovering the alternative of antimicrobial drugs for the treatment of infectious disease.

Antioxidant Capacity and Radical Scavenging Effect of Polyphenol Rich Mallotus philippenensis Fruit Extract on Human Erythrocytes: An In Vitro Study

Mallotus philippinensis is an important source of molecules with strong antioxidant activity widely used medicinal plant. Previous studies have highlighted their anticestodal, antibacterial, wound healing activities, and so forth. So, present investigation was designed to evaluate the total antioxidant activity and radical scavenging effect of 50% ethanol fruit glandular hair extract (MPE) and its role on Human Erythrocytes. MPE was tested for phytochemical test followed by its HPLC analysis. Standard antioxidant assays like DPPH, ABTS, hydroxyl, superoxide radical, nitric oxide, and lipid peroxidation assay were determined along with total phenolic and flavonoids content. Results showed that MPE contains the presence of various phytochemicals, with high total phenolic and flavonoid content. HPLC analysis showed the presence of rottlerin, a polyphenolic compound in a very rich quantity. MPE exhibits significant strong scavenging activity on DPPH and ABTS assay. Reducing power showed dose dependent increase in concentration absorption compared to standard, Quercetin. Superoxide, hydroxyl radical, lipid peroxidation, nitric oxide assay showed a comparable scavenging activity compared to its standard. Our finding further provides evidence that Mallotus fruit extract is a potential natural source of antioxidants which have a protective role on human Erythrocytes exhibiting minimum hemolytic activity and this justified its uses in folklore medicines.

In Vivo Healing Potential of Aegle marmelos in Excision, Incision, and Dead Space Wound Models

The study incorporates the wound healing potential of Aegle marmelos fruit pulp extract (AME) on excision, incision, and dead space wound models in rats. AME (200 mg/kg) was administered orally once daily for variable days depending on the type of wound ulcer study. AME was studied for its wound breaking strength (incision wound), rate of contraction, period of epithelization and histology of skin (excision model), and granulation tissue free radicals, antioxidants, acute inflammatory marker, and connective tissue markers and deep connective tissue histology (dead space wound). Complete wound contraction and epithelization were observed at the 20th day after treatment with AME as compared to the 24th day in control rats. Mean epithelization period and scar area were decreased while wound breaking strength was increased with AME compared with control. Granulation tissue showed increased levels of collagen determinants (33.7 to 64.4%, P < 0.001) and antioxidants (13.0 to 38.8%, P < 0.05 to P < 0.001), whereas markers of oxidative stress (55.0 to 55.6%, P < 0.001) and myeloperoxidase (21.3%, P < 0.001) were decreased in AME treated group. A. marmelos seems to promote wound healing by enhancing connective tissue formation and antioxidants status with decrease in free radicals and myeloperoxidase having tissue damaging effects.

Protective effects of Aegle marmelos fruit pulp on 2,4,6-trinitrobenzene sulfonic acid-induced experimental colitis.

Background: Aegle marmelos (AM) fruit has been advocated in indigenous system of medicine for the treatment of various gastrointestinal disorders, fever, asthma, inflammations, febrile delirium, acute bronchitis, snakebite, epilepsy, leprosy, myalgia, smallpox, leucoderma, mental illnesses, sores, swelling, thirst, thyroid disorders, tumours and upper respiratory tract infections. Objective: The objective of this study was to study the curative effect of 50% ethanol extract of dried fruit pulp of AM (AME) against 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis. Materials and Methods: AME (200 mg/kg) was administered orally, once daily for 14 days after TNBS-induced colitis. Rats were given intracolonic normal saline or TNBS alone or TNBS plus oral AME. AME was studied for its in vitro antibacterial activity against Gram-negative intestinal bacteria and on TNBS-induced changes in colonic damage, weight and adhesions (macroscopic and microscopic), diarrhea, body weight and colonic levels of free radicals (nitric oxide and lipid peroxidation), antioxidants (superoxide dismutase, catalase and reduced glutathione) and pro-inflammatory marker (myeloperoxidase [MPO]) in rats. Results: AME showed antibacterial activity against intestinal pathogens and decreased colonic mucosal damage and inflammation, diarrhea, colonic free radicals and MPO and enhanced body weight and colonic antioxidants level affected by TNBS. The effects of AME on the above parameters were comparable with sulfasalazine, a known colitis protective drug (100 mg/kg, oral). Conclusion: AME shows curative effects against TNBS-induced colitis by its antibacterial activity and promoting colonic antioxidants and reducing free radicals and MPO-induced colonic damage.

Pharmacological evaluation of extracts of Hedychium spicatum (Ham-ex-Smith) rhizome

Hedychium spicatum (Ham-ex-Smith), known as Shati in Ayurvedic classics, is documented for the treatment of cough, hiccough, fever and asthma. The present study includes the evaluation of aqueous and ethanolic extracts of the dried rhizome of H. spicatum for anti-histaminic and ulcer-protective activities in guinea pig (GP), anti-inflammatory and analgesic activities in rat and acute toxicity in mouse. The extracts were administered orally, daily as suspension, in 1% carboxymethyl cellulose either for 7 days in GP studies or 60 min before or just before experiment in rats and mice. An initial dose-dependent anti-histaminic action of both the extracts (100, 200 and 400 mg/kg) was performed against histamine-induced bronchospasm in GPs. The 200 mg/ kg dose of aqueous and ethanolic extracts was selected both in GP and rat for further studies. GPs treated with aqueous and ethanolic extracts showed gastric ulcer protection against histamine-induced gastric ulcer compared with the control group. Both the extracts also showed an anti-inflammatory effect against carrageenan-induced paw edema in rats from 1 h onwards, and this was maximum at 3 h. Analgesic effect was determined by using hot plate and tail flick tests in rats, and both the extracts at 200 mg/kg showed a significant increase in the latent period from 30 min onwards till 120 min of their study period. Both the extracts did not show any toxic effect like increased motor activity, salivation, clonic convulsion, coma and death in mice even at the 2000 mg/kg dose (nearly 10 times of the optimal effective dose), indicating the safety of the extracts. The result confirms the indigenous use of this plant in respiratory disorders.

Anti-infl ammatory activity of two classical formulations of Laghupanchamula in rats

Background: Laghupanchamula denotes combinations of roots of five herbs. However, in Ayurvedic classics besides four common herbs viz. Kantakari, Brihati, Shaliparni, and Prinshniparni, the fifth one is either Gokshura (Laghupanchamula with Gokshura LPG) or Eranda (Laghupanchamula with Eranda LPE), and both formulations have been documented to have shothahara (anti-inflammatory) action. Objectives: The present study was undertaken to compare the anti-inflammatory activity of 50% ethanolic extract of LPG (LPGE) and LPE (LPEE) in rats and safety in mice. Materials and Methods: LPGE and LPEE were given orally, administered either just before or 60 min before experiment on mice and for 7 days to rats. Paw edema was induced by carrageenan (acute) and formalin (sub-acute), whereas granuloma pouch (sub-acute) was induced by turpentine in rats. Results: Both LPGE and LPEE (1.0 g/kg) at 3 h after their administration showed inhibition of formalin-induced paw edema by 46.2% and 44.3% (P < 0.001) and carrageenan-induced paw edema by 53.9% and 60.4% (P < 0.001), respectively. After 7 days of treatment, both LPGE and LPEE showed 26.3% (P < 0.01) and 32.5% (P < 0.05) inhibition, respectively, against formalin-induced paw edema, and reduced weight of turpentine-induced granuloma pouch by 42.8% and 36.1% (P < 0.001), and volume of exudates by 31.2% and 36.2% (P < 0.001), respectively. No acute toxicity was observed in mice even with a 10.0-g/kg dose of both extracts. Conclusion: LPGE and LPEE significantly reduced acute and sub-acute inflammation, and showed effective and similar anti-inflammatory activity. They seemed to be safe, and use of both formulations in the Laghupanchamula for their anti-inflammatory activity is, thus, authenticated.

Effect of embelin on lithium–induced nephrogenic diabetes insipidus in albino rats

Objective To evaluate the nephroprotective and anti–polyuric role of embelin on lithium induced nephrogenic diabetes insipidus (NDI) in albino rats.

Effect of Azadirachta indica leaves extract on acetic acid-induced colitis in rats: Role of antioxidants, free radicals and myeloperoxidase

Abstract Objective To evaluate the healing effects of extract of dried leaves of Azadirachta indica (Neem) on acetic acid-induced colitis in rats. Neem tree is known as ‘arishtha’ in Sanskrit, meaning ‘reliever of sicknesses’. Methods 50% ethanolic extract of Azadirachta indica leaves was administered orally, once daily for 14 days in rats after the induction of colitis with acetic acid and 500 mg/kg dose of extract was found to have an optimal effect against acetic acid-induced colonic damage score, weight and adhesions (Macroscopic). Effect of Azadirachta indica extract was then further studied on various physical (mucous/blood in stool, food and water intake and body weight changes), colonic mucosal damage and inflammation (microscopic), antibacterial and biochemical parameters viz. i) antioxidants (superoxide dismutase, catalase and reduced glutathione) and ii) free radicals (nitric oxide and lipid peroxidation) and myeloperoxidase (acute inflammatory marker) activities in acetic acid-induced colitis. Results Azadirachta indica extract decreased colonic mucosal damage and inflammation (macroscopic and microscopic), mucous/bloody diarrhea, fecal frequency and increased body weight. Azadirachta indica extract showed intestinal antibacterial activity and enhanced the antioxidants but decreased free radicals and myeloperoxidase activities. Acute toxicity study indicated no mortality or other ANS or CNS related adverse effects even with 5.0 g/kg dose (10 times of effective dose) indicating its safety. Conclusions Azadirachta indica seemed to be safe and effective in colitis by its predominant effect on promoting antioxidant status and decreasing intestinal bacterial load, free radicals and myeloperoxidase responsible for tissue damage and delayed healing.

Isolation and Evaluation of Antimicrobial Activities of Endophytic Fungal Extract from Mallotus philippinensis Muell

Mallotus philippinensis Muell an important medicinal plant in the present study i.e., endophytic extracts of different fungi associated with asymptomatic were evaluated for antimicrobial activity. This will be a new promising source of antimicrobials known as endophytes. In this preliminary study, different parts of plants viz. steam and leaves were used for isolation of endophytic fungi and fermented. The endophytic fungi were identified based on their morphology and characteristics of fungal spores. The cell free ferment broth was subjected to antimicrobial assay against different human pathogenic microbes viz. Escherichia coli, Staphylococcus aureus, E. faecalis, K .pneumoniae and P. aeruginosa and three fungal pathogenic strains of candida i.e. C. albicans, C. tropicalis and C. krusie by using standard disc diffusion method. The results showed that some of these endophytic extracts had prominent antimicrobial inhibitory effects. Among these fungal endophytes, two strains Alternaria spp., Pestalotiopsis spp., and Phompsis spp. showed the strongest antimicrobial activities against P. aeruginosa, S. aureus and E. faecalis. An endophyte Pestalotiopsis spp. had the most pronounced effect on all the Candida strains exhibited the strongest antifungal activity. This present study has proven that both these medicinal plants may be a rich source of endophytic fungi with potential to produce bioactive producing antibacterial and antifungal activities. These antipathogenic endophytes could be applied as new sources of antibiotics in agriculture and pharmaceutical industry.